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Updated: Feb 3, 2026
05:53
Trans-inner Cell Mass Injection of Embryonic Stem Cells Leads to Higher Chimerism Rates
Published on: May 29, 2018
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Chimeric Mouse Generation by ES Cell Blastocyst Microinjection and Uterine Transfer
Yubin Du1, Wen Xie1, Fan Zhang1
1Transgenic Core Facility, Division of Intramural Research, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Methods in Molecular Biology (Clifton, N.J.)
|October 25, 2018
Summary
Generating chimeric mice using embryonic stem (ES) cells and blastocyst microinjection remains a valuable technique for creating sophisticated genetic models and advancing stem cell research.
Area of Science:
- * Stem Cell Biology
- * Developmental Biology
- * Genetic Engineering
Background:
- * Conventional embryonic stem (ES) cell-mediated knockout technology relies on generating chimeric mice via blastocyst microinjection.
- * While CRISPR technology has reduced the need for microinjection, it remains essential for specific applications.
- * This technique is crucial for creating complex genetic models and for fundamental stem cell research.
Purpose of the Study:
- * To provide a detailed protocol for generating chimeric mice using ES cells.
- * To outline the essential steps from cell culture to surrogate mother implantation.
- * To serve as a guide for researchers in stem cell and genetic engineering fields.
Main Methods:
- * Derivation and inactivation of Mouse Embryonic Fibroblast (MEF) feeder cells.
- * Culture and handling of mouse ES cells.
- * Collection and microinjection of blastocysts.
- * Implantation of microinjected blastocysts into pseudopregnant surrogate mothers.
Main Results:
- * Successful generation of chimeric mice using the described ES cell microinjection protocol.
- * Establishment of a reproducible method for producing genetically modified mice.
- * Validation of the technique for creating advanced genetic models.
Conclusions:
- * Blastocyst microinjection with ES cells is a robust and valuable method for generating chimeric mice.
- * This technique continues to be indispensable for sophisticated genetic engineering and stem cell research.
- * The detailed protocol facilitates the production of custom genetic models for various research applications.
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